Structure of SARS Coronavirus Spike Glycoprotein

SARS 冠状病毒刺突糖蛋白的结构

• 批准号: 6758154
• 负责人: MICHAEL JEFFREY ROOT
• 金额: $ 23.4万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6758154
• 关键词: SARS virus; X ray crystallography; antiviral agents; biophysics; cell line; conformation; drug discovery /isolation; emerging infectious disease; glycoproteins; host organism interaction; mass spectrometry; peptide chemical synthesis; protein structure; protein structure function; proteolysis; recombinant proteins; severe acute respiratory syndrome; structural biology; transfection; virus infection mechanism; virus protein

DESCRIPTION (provided by applicant): Severe acute respiratory syndrome (SARS) has recently emerged as a new human disease with a mortality rate as high as 15%. The genome of the putative etiological agent, SARS-associated coronavirus (SCV), contains a spike glycoprotein that is implicated in viral attachment and entry by homology to similar proteins in related coronaviruses. These spike glycoproteins contain two regions of hydrophobic heptad repeat sequences characteristic of the viral trimer-of-hairpins structural motif. Formation of a trimer-of-hairpins is an essential step in the membrane fusion process of other enveloped viruses, including HIV-1, Ebola virus and respiratory syncytial virus. Moreover, for these unrelated viruses, polypeptides and small molecules that disrupt the formation of this crucial structure are potent inhibitors of viral infectivity. The studies proposed here test the trimer-of-hairpins structural hypothesis for SCV and develop membrane fusion assays to test possible inhibitors of SCV spike glycoprotein. The specific aims are: 1) To generate and biophysically characterize polypeptides derived from the heptad repeat regions of SCV spike glycoprotein, and to solve the structure of particularly stable constructs using x-ray crystallography; and 2) To develop a cell-cell fusion assay and/or a pseudoviral infectivity assay to assess the fusion activity of the SCV spike glycoprotein, and to test polypeptides derived from the heptad repeat regions for potential inhibitory activity. These experiments will explore the structure and function of the SCV spike glycoprotein, essential information for the future development of antiviral therapeutics and vaccines.

描述（申请人提供）： 严重急性呼吸系统综合症（SARS）是最近出现的一种新的人类疾病，死亡率高达15%。假定的病原体 SARS 相关冠状病毒 (SCV) 的基因组含有一种刺突糖蛋白，该糖蛋白与相关冠状病毒中的类似蛋白具有同源性，与病毒的附着和进入有关。这些刺突糖蛋白含有两个疏水性七肽重复序列区域，是病毒发夹三聚体结构基序的特征。发夹三聚体的形成是其他包膜病毒（包括 HIV-1、埃博拉病毒和呼吸道合胞病毒）膜融合过程中的重要步骤。此外，对于这些不相关的病毒来说，破坏这一关键结构形成的多肽和小分子是病毒感染性的有效抑制剂。这里提出的研究测试了 SCV 的发夹三聚体结构假说，并开发了膜融合测定法来测试 SCV 刺突糖蛋白的可能抑制剂。具体目标是： 1) 生成源自 SCV 刺突糖蛋白七肽重复区域的多肽并对其进行生物物理表征，并使用 X 射线晶体学解析特别稳定的构建体的结构； 2)开发细胞-细胞融合测定和/或假病毒感染性测定，以评估SCV刺突糖蛋白的融合活性，并测试源自七肽重复区域的多肽的潜在抑制活性。这些实验将探索 SCV 刺突糖蛋白的结构和功能，这是未来抗病毒疗法和疫苗开发的重要信息。